摘要
目的探讨欧前胡素(IMP)对急性髓系白血病(AML)细胞恶性生物学行为的影响以及对MAPK/mTOR/p70S6K信号通路的调节机制。方法将人AML细胞HL-60进行培养传代,并分为对照组(未处理组),ERK抑制剂组(PD98059组),IMP低、中、高剂量组,IMP高剂量+ERK激活剂组(IMP高+Cearoin组),每组均设置6个重复。MTT法检测IMP对HL-60细胞的毒性;MTT法、软琼脂克隆形成实验检测HL-60细胞的增殖活性;流式细胞仪术检测HL-60细胞凋亡;Transwell小室检测HL-60细胞迁移和侵袭能力;ELISA检测HL-60细胞培养液中TNF-α、IL-1β、IL-6的水平;Western blot检测通路相关蛋白及Bcl-2、Beclin-1、LC3Ⅱ/Ⅰ蛋白的表达。结果与未处理组比较,PD98059组和IMP低、中、高剂量组HL-60细胞的存活率、克隆形成数量、迁移和侵袭数量、促炎因子TNF-α、IL-1β、IL-6水平以及ERK1/2、mTOR、p70S6K磷酸化水平、Bcl-2表达显著降低(P<0.05),细胞凋亡率、自噬相关蛋白Beclin-1、LC3Ⅱ/Ⅰ显著升高(P<0.05);与IMP高剂量组比较,IMP高+Cearoin组中ERK激活剂明显消除了IMP对上述指标的影响(P<0.05)。结论IMP可能通过抑制MAPK/mTOR/p70S6K信号通路,抑制AML细胞增殖、迁移和侵袭,促进细胞自噬和凋亡。
Objective To investigate the influences of imperatorin(IMP)on the malignant biological behaviors of acute myeloid leukemia(AML)cells via mediating the MAPK/mTOR/p70S6K signaling pathway.Methods The human AML cell line HL-60 was cultured and passaged.HL-60 cells were treated with blank control(control group),ERK inhibitor PD98059(PD98059 group),low-dose IMP(low-dose IMP group),medium-dose IMP(medium-dose IMP group),high-dose IMP(high-dose IMP group)and high-dose IMP plus ERK activator Cearoin(high-dose IMP+Cearoin group),with 6 replicates per group.The toxicity of IMP to HL-60 cells was detected by MTT assay.The proliferation of HL-60 cells was detected by MTT assay and soft agar colony formation assay.Cell apoptosis was detected by flow cytometry.Cell migration and invasion were detected by Transwell assay.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of TNF-α,IL-1βand IL-6 in the culture medium of HL-60 cells.Western blot was performed to detect protein expressions of key factors in the MAPK/mTOR/p70S6K signaling pathway,Bcl-2,Beclin-1,and LC3Ⅱ/Ⅰ.Results Compared with those of the control group,significantly lower survival,number of colonies,and migratory and invasive cells,relative levels of proinflammatory factors(TNF-α,IL-1β,IL-6),p-ERK1/2,p-mTOR,p-p70S6K and Bcl-2,and significantly higher apoptotic rate,and expression levels of autophagy proteins(Beclin-1,LC3Ⅱ/Ⅰ)were detected in PD98059 group and low-dose,medium-dose and high-dose IMP group(P<0.05).Compared with those of high-dose IMP group,the regulatory effects of IMP on the abovementioned indicators were reversed by the ERK activator Cearoin(P<0.05).Conclusion IMP inhibits the proliferation,migration and invasion of AML cells and promotes autophagy and apoptosis by inhibiting the MAPK/mTOR/p70S6K signaling pathway.
作者
李慧
张吴霞
熊烨
LI Hui;ZHANG Wuxia;XIONG Ye(Laminar Flow Research Ward of Clinical Trial Center,West China Hospital of Sichuan University,Sichuan,Chengdu 610044,China;不详)
出处
《河北医药》
CAS
2023年第13期1941-1945,共5页
Hebei Medical Journal
基金
四川省自然科学基金(编号:2022NSFSC1463)。
